+ Site Statistics
+ Search Articles
+ Subscribe to Site Feeds
EurekaMag Most Shared ContentMost Shared
EurekaMag PDF Full Text ContentPDF Full Text
+ PDF Full Text
Request PDF Full TextRequest PDF Full Text
+ Follow Us
Follow on FacebookFollow on Facebook
Follow on TwitterFollow on Twitter
Follow on LinkedInFollow on LinkedIn

+ Translate

Multi-site evaluation of CD34+ enumeration methods with processed cord blood

Multi-site evaluation of CD34+ enumeration methods with processed cord blood

Blood 102(11): 460a, November 16

Umbilical Cord Blood (UCB) is collected and processed as a source of hematopoietic stem cells (HPC) for allogeneic transplantation. HPC enumeration in UCB presents unique challenges due to the heterogeneous cell populations that include nucleated red blood cells, epithelial cells and multiple CD34+ populations, some of which are not HPC. Most studies have shown that sequential gating (ISHAGE procedure) combined with reference counting beads allows the highest degree of correlation between centers when the specimen is peripheral blood or apheresis product. This procedure has now been incorporated into two commercial methods, the Stem-KitTM procedure (BC, Beckman-Coulter) and the TruCOUNTTM procedure (BD, BD Biosciences). Standardization of procedures for CD34+ HPC enumeration would be beneficial for improvement of inter-laboratory comparisons. We conducted two studies to compare identical processed samples to assess inter-laboratory comparability. In study 1, four UCB processing laboratories processed 25 UCB (HES-sedimentation procedure) for cryopreservation and determined CD34+ HPC number using their routine procedure prior to cryopreservation. Identical aliquots were shipped to a central laboratory by overnight courier at 1-10degreeC for testing using the two commercially available methods. In study 2, two sites performed CD34+ HPC enumeration on 25 duplicate UCB specimens using identical commercial procedures (BC and BD). Manufacturer's instructions were followed without deviation. Linear regression analysis and the paired Student's t test were used as tests of significance. In study 1, the central lab showed good correlation between the BC and the BD methods (r=0.94) for absolute CD34+ HPC numbers. The CD34+ HPC numbers obtained by 2 out of 4 testing sites were comparable to the those obtained by the central laboratory (BC method, r=0.99, 0.81). The two other laboratories showed poor correlation with one laboratory actually showing a statistical significance in deviation from the central laboratory (pltoreq0.05 and r=0.62, 0.59 for BC method). Factors that contributed to the differences between labs were: determination of the nucleated cell counts; type of hematology analyzer used for use in dual platform methods (hematology analyzer vs particle counter); inclusion of nucleated red blood cells in the NC count; and fixing of samples prior to CD34+ HPC enumeration. In study 2 the central laboratory and a research laboratory determined comparable CD34+ cell levels with the BC and BD methods (r=0.96, 0.93 respectively). These data indicate that the BC and BD methods can be utilized to measure CD34+ cell levels in processed cord blood samples. CD34+ cell counts between laboratories can be comparable, particularly when single platform methods were used.

(PDF 0-2 workdays service: $29.90)

Accession: 035349515

Download citation: RISBibTeXText

Related references

Multi-site evaluation of the BD Stem Cell Enumeration Kit for CD34(+) cell enumeration on the BD FACSCanto II and BD FACSCalibur flow cytometers. CytoTherapy 16(11): 1558-1574, 2015

Comparison of four methods for CD34+ cell enumeration in cord blood. European Journal of Histochemistry 41 Suppl 2: 37-38, 1997

Enumeration of viable CD34(+) cells by flow cytometry in blood, bone marrow and cord blood: results of a study of the novel BD™ stem cell enumeration kit. CytoTherapy 13(4): 449-458, 2011

Enumeration of viable CD34 cells in cord blood and cord blood products. Blood 90(10 SUPPL 1 PART 2): 342B-343B, Nov 15, 1997

The enumeration of CD34+ progenitor cells in cord blood and in peripheral blood after G-CSF mobilisation Reproducibility and stability. Experimental Hematology (Charlottesville) 24(9): 1032, 1996

A comparison of cryopreservation methods: Slow-cooling vs. rapid-cooling based on cell viability, oxidative stress, apoptosis, and CD34+ enumeration of human umbilical cord blood mononucleated cells. Bmc Research Notes 4(): 371-371, 2011

Analysis of variance in CD34+ progenitor cell enumeration of cord blood. British Journal of Haematology 102(1): 158, July 1, 1998

Lack of standardization of methodology for enumeration of CD34 in banked umbilical cord blood. Blood 98(11 Part 2): 333b, November 16, 2001

Sensitive detection and enumeration of CD34+ cells in peripheral and cord blood by flow cytometry. Experimental Hematology 22(10): 1003-1010, 1994

Enumeration of CD34-positive stem cells: evaluation and comparison of three methods. Journal of HematoTherapy 6(3): 213-226, 1997

CD34+ cell selection of umbilical cord blood samples previously processed with hydroxyethyl starch. Blood 92(10 SUPPL 1 PART 1-2): 314B, Nov 15, 1998

Enumeration of CD34+ cells in cord blood: A variation on a single-platform flow cytometric method based on the ISHAGE gating strategy. Cytometry 46(4): 254-261, August 15, 2001

Direct comparison of 3 analytical methods for enumeration of CD34+ cells in peripheral blood and leukapheresis products. Bone Marrow Transplantation 0(PROGRAM ABSTR ): 109, 1994

Evaluation of new rapid methods used for enumeration of aerobic bacteria in processed meats and environmental samples. Abstracts of the General Meeting of the American Society for Microbiology 99: 524, 1999